JP2009536211A - MGLUR5 Modulator III - Google Patents

Abstract

  The present invention relates to a novel compound, a process for its preparation, its use in therapy and a pharmaceutical composition comprising the novel compound.

Description

  The present invention relates to novel compounds, their use in therapy and pharmaceutical compositions comprising said novel compounds.

  Glutamate is the major excitatory neurotransmitter in the mammalian central nervous system (CNS). Glutamate binds to and activates cell surface receptors, thereby producing its effects in neurons of the central nervous system. These receptors are based on the structural features of the receptor protein, the means by which the receptor transmits signals to the cell, and the pharmacological profile of two major types, ion channel and metabotropic glutamate receptors. It is divided into bodies.

Metabotropic glutamate receptor (mGluR) is a G protein-coupled receptor that activates various intracellular second messenger systems after binding of glutamate. Activation of mGluR in intact mammalian neurons elicits one or more of the following responses: activation of phospholipase C; increase in phosphoinositide (PI) hydrolysis; intracellular calcium release; phospholipase Activation of D; activation or inhibition of adenyl cyclase; increase or decrease in formation of cyclic adenosine monophosphate (cAMP); activation of guanylyl cyclase; increase in formation of cyclic guanosine monophosphate (cGMP); phospholipase Activation of A ₂ ; increase in arachidonic acid release; and increase or decrease in the activity of voltage and ligand-gated ion channels. Schoepp et al., Trends Pharmacol. Sci. 14: 13 (1993), Schoepp, Neurochem. Int. 24: 439 (1994), Pin et al., Neuropharmacology 34: 1 (1995), Bordi and Ugolini, Prog. Neurobiol 59:55 (1999).

  In molecular cloning, eight distinct mGluR subtypes, designated mGluR1-mGluR8, have been identified. Nakanishi, Neuron 13: 1031 (1994), Pin et al., Neuropharmacology 34: 1 (1995), Knopfel et al., J. Med. Chem. 38: 1417 (1995). There is further receptor diversity. It arises through the expression of alternatively spliced forms of the species mGluR subtype. Pin et al., PNAS 89: 10331 (1992), Minakami et al., BBRC 199: 1136 (1994), Joly et al., J. Neurosci. 15: 3970 (1995).

  Metabotropic glutamate receptor subtypes are classified into three groups, Group I, Group II and Group III mGluRs based on amino acid sequence homology, the second messenger system utilized by the receptor, and its pharmacological properties. It can be further divided. Group I mGluRs include mGluR1, mGluR5 and their alternatively spliced variants. Binding of agonists to these receptors results in activation of phospholipase C and subsequent mobilization of intracellular calcium.

Neurological, psychiatric and pain disorders Attempts to elucidate the physiological role of Group I mGluR suggest that activation of this receptor induces neuronal excitation. Various studies have shown that group I mGluR agonists can produce post-synaptic excitation when applied to neurons not only in the hippocampus, cerebral cortex, cerebellum and thalamus but also in other CNS regions. Evidence has shown that this excitation is due to direct activation of post-synaptic mGluR, but it also suggests that activation of pre-synaptic mGluR occurs and neurotransmitter release is enhanced. Baskys, Trends Pharmacol. Sci. 15: 92 (1992), Schoepp, Neurochem. Int. 24: 439 (1994), Pin et al., Neuropharmacology 34: 1 (1995), Watkins et al., Trends Pharmacol. Sci. 15:33 (1994).

  Metabotropic glutamate receptors are involved in many normal processes in the mammalian CNS. Activation of mGluR has been shown to be required for induction of hippocampal long-term potentiation and cerebellar long-term depression. Bashir et al., Nature 363: 347 (1993), Bortolotto et al., Nature 368: 740 (1994), Aiba et al., Cell 79: 365 (1994), Aiba et al., Cell 79: 377 (1994 Meller et al., Neuroreport 4: 879 (1993), Bordi and Ugolini, Brain Res. 871: 223 (1999) have also shown the role of mGluR activation in nociception and analgesia. In addition, mGluR activation is associated with a variety of other, including synaptic transmission, neuronal development, apoptotic neuronal death, synaptic plasticity, spatial learning, olfactory memory, central control of cardiac activity, arousal, motor regulation and control of vestibular eye reflexes. It has been suggested to play a regulatory role in the normal process. Nakanishi, Neuron 13: 1031 (1994), Pin et al., Neuropharmacology 34: 1, Knopfel et al., J. Med. Chem. 38: 1417 (1995).

  Furthermore, group I metabotropic glutamate receptors, and in particular mGluR5, have been suggested to play a role in disorders affecting various pathophysiological processes and the CNS. These include stroke, head trauma, hypoxia and ischemic injury, hypoglycemia, epilepsy, neurodegenerative disorders such as Alzheimer's disease, and pain. Schoepp et al., Trends Pharmacol. Sci. 14: 13 (1993), Cunningham et al., Life Sci. 54: 135 (1994), Hollman et al., Ann. Rev. Neurosci. 17:31 (1994), Pin et al., Neuropharmacology 34: 1 (1995), Knopfel et al., J. Med. Chem. 38: 1417 (1995), Spooren et al., Trends Pharmacol. Sci. 22: 331 (2001), Gasparini et al. Curr. Opin. Pharmacol. 2:43 (2002), Neugebauer Pain 98: 1 (2002). Many of these pathologies are thought to be due to excess glutamate-induced excitement of CNS neurons. It is done. Since Group I mGluRs are thought to enhance glutamate-mediated neuronal excitement via post-synaptic mechanisms and enhanced presynaptic glutamate release, their activation is probably responsible for the pathology. Accordingly, selective antagonists of Group I mGluR receptors may be therapeutically beneficial, particularly as neuroprotective agents, analgesics or anticonvulsants.

  In general, recent advances in the elucidation of metabotropic glutamate receptors, and in particular the neurophysiological role of group I, make these receptors in the treatment of acute and chronic neurological and psychiatric disorders and chronic and acute pain disorders. It has been established as a promising drug target in the future.

Gastrointestinal disorders The lower esophageal sphincter (LES) tends to relax intermittently. As a result, the mechanical barrier may be temporarily lost in such a case, and the fluid from the stomach may move to the esophagus, and the event is hereinafter referred to as “reflux”.

  Gastroesophageal reflux disease (GERD) is the most common upper gastrointestinal disease. Current drug therapy aims at reducing gastric acid secretion or neutralizing acid in the esophagus. The main mechanism behind reflux is believed to be the hypotonic lower esophageal sphincter. However, for example Holloway & Dent (1990) Gastroenterol. Clin. N. Amer. 19, pp. 517-535, most reflux episodes are induced by transient lower esophageal sphincter relaxation (TLESR), ie swallowing. But not during relaxation. In addition, gastric acid secretion is usually found to be normal in patients with GERD.

  The novel compounds of the present invention are believed to be useful for the inhibition of transient lower esophageal sphincter relaxation (TLESR) and thus for the treatment of gastroesophageal reflux disorder (GERD).

It is well known that certain compounds can produce undesirable effects on human cardiac repolarization, which is observed as an extension of the QT interval on the electrocardiogram (ECG). In extreme situations, this drug-induced prolongation of the QT interval is described by Torsade de Pointe (TdP; Vandenberg et al. HERG K ⁺ channels: friend and foe. Trends Pharmacol Sci 2001; 22: 240-246) It leads to a type of cardiac arrhythmia that can eventually lead to ventricular fibrillation and sudden death. The primary event in this syndrome is the inhibition of the fast component of the delayed rectifier potassium current (IKr) by these compounds. The compound encodes a human ether-a-go-go-related gene (hERG), a channel protein opening alpha subunit that carries this current subunit (Aperture-forming alpha sub-units)

  Since IKr plays an important role in the repolarization of the cardiac action potential, its inhibition delays the repolarization and this appears as an extension of the QT interval. Prolonging the QT interval is not a safety concern in itself, but it carries the risk of cardiovascular side effects and in a small percentage of people can lead to TdP and degeneration to ventricular fibrillation.

  In general, the compounds of the invention have low activity against the potassium channel encoding hERG. In this regard, low in vitro activity against hERG indicates low in vivo activity.

  Also, in order to increase the effectiveness of the drug, it is desirable that the drug has good metabolic stability. In vitro stability to metabolism of human microsomes indicates in vivo stability to metabolism.

  New potent mGluR agonists and antagonists that show selectivity for mGluR subtypes, particularly Group I receptor subtypes, most specifically mGluR5, are needed because of their physiological and pathophysiological significance. ing.

  An object of the present invention is to provide a compound that exhibits activity at a metabotropic glutamate receptor (mGluR), particularly at the mGluR5 receptor. In particular, the compounds of the invention act primarily on the periphery, ie have a limited ability to cross the blood brain barrier.

（式中、
Ｒ¹は、メチル、ハロゲン又はシアノであり；
Ｒ²は、水素又はフルオロであり；
Ｒ³は、水素、フルオロ又はＣ₁−Ｃ₃アルキルであり；
Ｒ⁴は、Ｃ₁−Ｃ₃アルキル又はシクロプロピルであり；
Ｘは、

であり；
そしてＺは、

であり；
ここにおいて、
Ｒ⁵は、水素、Ｃ₁−Ｃ₃アルキル、Ｃ₁−Ｃ₃ハロアルキル、Ｃ₁−Ｃ₃アルコキシ、Ｃ₁−Ｃ₃ハロアルコキシ；又はハロゲンであり；
Ｒ⁶は、水素、Ｃ₁−Ｃ₃アルキル、Ｃ₁−Ｃ₃ハロアルキル、Ｃ₁−Ｃ₃アルコキシ；Ｃ₁−Ｃ₃ハロアルコキシ；又はハロゲンであり；
Ｒ⁷は、水素、フルオロ又はＣ₁−Ｃ₃アルキルである）
並びにその医薬上許容しうる塩、水和物、アイソフォーム、互変異性体及び／又は鏡像異性体に関する。 The present invention relates to a compound of formula I:

(Where
R ¹ is methyl, halogen or cyano;
R ² is hydrogen or fluoro;
R ³ is hydrogen, fluoro or C ₁ -C ₃ alkyl;
R ⁴ is C ₁ -C ₃ alkyl or cyclopropyl;
X is

Is;
And Z is

Is;
put it here,
R ⁵ is hydrogen, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, C ₁ -C ₃ alkoxy, C ₁ -C ₃ haloalkoxy; or halogen;
R ⁶ is hydrogen, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, C ₁ -C ₃ alkoxy; C ₁ -C ₃ haloalkoxy; or halogen;
R ⁷ is hydrogen, fluoro or C ₁ -C ₃ alkyl)
As well as pharmaceutically acceptable salts, hydrates, isoforms, tautomers and / or enantiomers thereof.

In one embodiment, R ¹ is halogen or cyano.
In a further embodiment, R ¹ is chloro. In a further embodiment, R ¹ is cyano.
In a further embodiment, R ² is hydrogen.
In a further embodiment, R ³ is hydrogen or fluoro.
In a further embodiment, R ⁴ is C ₁ -C ₂ alkyl.
In a further embodiment, R ⁴ is methyl.
In a further embodiment, R ⁵ is hydrogen, C ₁ -C ₂ alkyl or C ₁ -C ₂ alkoxy.
In a further embodiment, R ⁶ is hydrogen, C ₁ -C ₂ alkyl or C ₁ -C ₂ alkoxy.
In a further embodiment, R ⁷ is hydrogen or fluoro.

  Another embodiment is a pharmaceutical composition comprising a therapeutically effective amount of a compound of formula I as an active ingredient together with one or more pharmaceutically acceptable excipients, additives and / or inert carriers.

  Another embodiment described in more detail below relates to compounds of formula I for use in the manufacture of a medicament for treating a disorder mediated by mGluR5 in the treatment of a disorder mediated by mGluR5.

  Yet another embodiment relates to a method of treating a disorder mediated by mGluR5 comprising administering to a mammal a therapeutically effective amount of a compound of formula I.

  In another embodiment, there is provided a method of inhibiting mGluR5 receptor activation comprising treating a cell comprising said receptor with an effective amount of a compound of formula I.

  The compounds of the present invention are useful for therapy, particularly for the treatment of neurological, psychiatric, pain and gastrointestinal disorders.

  It will also be appreciated by those skilled in the art that certain compounds of the present invention can exist in solvated forms as well as unsolvated forms, eg, hydrated forms. It is further understood that the present invention encompasses all such solvated forms of the compounds of formula I.

  Also within the scope of the invention are salts of the compounds of formula I. In general, pharmaceutically acceptable salts of the compounds of the invention can be prepared using standard methods well known in the art, for example, a sufficiently basic compound such as an alkylamine with a suitable acid such as HCl, It is obtained by reacting with acetic acid or methanesulfonic acid to form a salt with a physiologically acceptable anion. Alternatively, a compound of the invention having a suitable acidic proton, such as a carboxylic acid or phenol, can be converted to 1 equivalent of an alkali metal or alkaline earth metal hydroxide or alkoxide (eg ethoxide or methoxide), or suitably Treatment with a basic organic amine (eg choline or meglumine) followed by production of the corresponding alkali metal (eg sodium, potassium or lithium) or alkaline earth metal (eg calcium) salt by conventional purification techniques . Furthermore, quaternary ammonium salts can be prepared by adding an alkylating agent, for example to a neutral amine.

  In one embodiment of the invention, the compound of the formula I is a pharmaceutically acceptable salt or solvate thereof, in particular an acid addition salt, such as hydrochloride, hydrobromide, phosphate, acetate, fumaric acid It can be converted to a salt, maleate, tartaric acid, citrate, methanesulfonate or p-toluenesulfonate.

The general terms used in the definition of formula I have the following meanings:
The halogen used herein is selected from chlorine, fluorine, bromine or iodine.

C ₁ -C ₃ alkyl is a straight or branched alkyl group having 1 to 3 carbon atoms, for example methyl, ethyl, n-propyl or isopropyl.

C ₁ -C ₃ alkoxy is an alkoxy group having 1 to 3 carbon atoms, for example methoxy, ethoxy, isopropoxy or n-propoxy.

C ₁ -C ₃ haloalkoxy is an alkoxy group having 1 to 3 carbon atoms in which at least one carbon atom has been replaced by a halogen atom, for example methoxy, ethoxy or n-propoxy.

  All chemical names were generated using software known as AutoNom accessed through ISIS draw.

  In Formula I above, X can exist in either of two possible orientations.

Pharmaceutical Compositions The compounds of the present invention can be formulated into conventional pharmaceutical compositions comprising a compound of formula I, or a pharmaceutically acceptable salt or solvate thereof, together with a pharmaceutically acceptable carrier or additive. Pharmaceutically acceptable carriers can be either solid or liquid. Solid formulations include, but are not limited to, powders, tablets, dispersible granules, capsules, cachets and suppositories.

  A solid carrier can be one or more substances which may also act as excipients, flavoring agents, solubilizers, lubricants, suspending agents, binders or tablet disintegrating agents. The solid carrier can also be an encapsulating material.

  In powders, the carrier is a finely divided solid, which is in a mixture with the finely divided compound of the invention, or the active component. In tablets, the active ingredient is mixed with the carrier having the necessary binding properties in suitable proportions and formed into the desired shape and size.

  For preparing suppository compositions, a low-melting wax such as a mixture of fatty acid glycerides and cocoa butter is first melted and the active ingredient is dispersed therein by, for example, stirring. The molten homogeneous mixture is then poured into convenient sized molds and allowed to cool and solidify.

  Suitable carriers include, but are not limited to, magnesium carbonate, magnesium stearate, talc, lactose, sugar, pectin, dextrin, starch, tragacanth, methylcellulose, sodium carboxymethylcellulose, a low melting wax, cocoa butter, and the like. I don't mean.

  The term composition is also intended to include the formulation of the active ingredient with an encapsulating material as the carrier for supplying the capsule, in which the active ingredient is surrounded by the carrier (with or without other carriers). So that the carrier is associated with the active ingredient. Similarly, cachets are included.

  Tablets, powders, cachets, and capsules can be used as solid dosage forms suitable for oral administration.

  Liquid compositions include solutions, suspensions and emulsions. For example, sterile water or aqueous propylene glycol solutions of the active compounds can be liquid formulations suitable for parenteral administration. Liquid compositions can also be formulated in solution in an aqueous polyethylene glycol solution.

  Aqueous solutions for oral administration can be prepared by dissolving the active component in water and adding suitable colorants, flavoring agents, stabilizers, and thickening agents as desired. Aqueous suspensions for oral use make the finely divided active ingredient a viscous material, such as natural synthetic rubber, resin, methylcellulose, sodium carboxymethylcellulose, and other suspending agents known in the pharmaceutical formulating art And can be produced by dispersing in water. A typical composition intended for oral use may contain one or more colorants, sweeteners, flavoring agents and / or preservatives.

  Depending on the mode of administration, the pharmaceutical composition comprises from about 0.05% w (mass%) to about 99% w of the compound of the invention, or comprises from about 0.10% w to 50% w, all masses The percentage is based on the total weight of the composition.

  The therapeutically effective amount for practicing the present invention can be determined by those skilled in the art using known criteria, including the age, weight and response of the individual patient, and for the disease to be treated or prevented. Interpreted in a frame.

Medical Use The compounds of the present invention are useful for treating conditions associated with excitatory activation of mGluR5 and inhibiting neuronal damage caused by excitatory activation of mGluR5. The compounds can be used to provide mGluR5 inhibitory action in mammals, including humans.

  Group I mGluR receptors, including mGluR5, are highly expressed in the central and peripheral nervous system and other tissues. Accordingly, the compounds of the present invention are expected to be well suited for the treatment of disorders mediated by mGluR5, such as acute and chronic neurological and psychiatric disorders, gastrointestinal disorders and chronic and acute pain disorders .

  The present invention relates to a compound of formula I as defined above for use in therapy.

  The present invention relates to compounds of formula I as defined above for use in the treatment of disorders mediated by mGluR5.

  The present invention relates to senile dementia of Alzheimer's disease, AIDS-induced dementia, Parkinson's disease, amyotrophic lateral sclerosis, Huntington's chorea, migraine, epilepsy, schizophrenia, depression, anxiety, acute anxiety, ophthalmic Disorders such as retinopathy, diabetic retinopathy, glaucoma, auditory neuropathic disorders such as tinnitus, chemotherapy-induced neuropathy, postherpetic neuralgia and trigeminal neuralgia, tolerance, dependence, fragile X, autism Relates to a compound of formula I as defined above for use in the treatment of symptom, mental retardation, schizophrenia and Down's syndrome.

  The present invention relates to migraine, inflammatory pain, neuropathic pain disorders such as diabetic neuropathy, arthritis and rheumatoid diseases, low back pain, postoperative pain and cancer, angina, renal or gallstone colic, menstruation, migraine and It relates to compounds of formula I as defined above for use in the treatment of pain associated with various conditions including gout.

  The present invention relates to compounds of formula I as defined above for use in the treatment of stroke, head trauma, hypoxia and ischemic injury, hypoglycemia, cardiovascular disease and epilepsy.

  The invention also relates to the use of a compound of formula I as defined above in the manufacture of a medicament for treating a disorder mediated by the mGluR group I receptor and any of the disorders mentioned above.

  One embodiment of the invention relates to the use of a compound of formula I in the treatment of gastrointestinal disorders.

  Another embodiment of the present invention is the inhibition of transient lower esophageal sphincter relaxation, the treatment of GERD, the prevention of gastroesophageal reflux, the treatment of discharge, the treatment of asthma, the treatment of laryngitis, the treatment of lung disease, the treatment of growth disorders It relates to the use of a compound of formula I for the manufacture of a medicament for the management, treatment of irritable bowel disease (IBS) and functional dyspepsia (FD).

  Another embodiment of this invention is directed to the use of a compound of formula I for treating overactive bladder or urinary incontinence.

  The term “TLESR”, transient lower esophageal sphincter relaxation, is referred to herein as Mittal, RK, Holloway, RH, Penagini, R., Blackshaw, LA, Dent, J., 1995; Transient lower esophageal sphincter relaxation. Gastroenterology 109, pp. 601-610.

  The term “reflux” is defined herein as that fluid from the stomach can enter the esophagus, at which time the mechanical barrier is temporarily lost.

  The term “GERD”, gastroesophageal reflux disease, is defined herein according to van Heerwarden, M.A., Smout A.J.P.M., 2000; Diagnosis of reflux disease. Bailliere's Clin. Gastroenterol. 14, pp. 759-774.

  The compounds of formula I above are obesity or overweight (eg, promoting weight loss and maintaining weight loss), preventing or reversing weight gain (eg, rebound, induced by medication or after smoking cessation), appetite and / or Or treat or prevent eating disorders (eg, bulimia, anorexia, bulimia and obsessive) and cravings (drugs, tobacco, alcohol, all appetizing macronutrients or non-essential foods) to adjust satiety Useful for.

  The invention also suffers from or is at risk for a mGluR5-mediated disorder and any of the disorders described above, comprising administering to a patient an effective amount of a compound of formula I as defined above. Provided are methods for treating disorders mediated by mGluR5 and any of the above disorders in a patient.

  The dose required for the therapeutic or prophylactic treatment of a particular disorder will necessarily vary depending on the host treated, the route of administration and the severity of the illness being treated.

  For the purposes of this specification, the terms “therapy” and “treatment” include prevention or prophylaxis unless specifically indicated to the contrary. Accordingly, the terms “therapeutic” and “therapeutically” should be construed accordingly.

  In this specification, unless stated otherwise, the terms “antagonist” and “inhibitor” shall mean a compound that, by any means, partially or completely blocks a transduction pathway that produces a response by a ligand. .

  The term “disorder”, unless stated otherwise, means all conditions and diseases associated with metabotropic glutamate receptor activity.

  One embodiment of the invention is a combination of a compound of formula I and an acid secretion inhibitor. The “combination” of the present invention can exist as a “fix combination” or a “kit of parts combination”. “Fixed combination” is defined as a combination in which (i) at least one acid secretion inhibitor; and (ii) at least one compound of formula I is present in one unit. A “part combination kit” is defined as a combination in which (i) at least one acid secretion inhibitor; and (ii) at least one compound of formula I is present in a plurality of units. The components of the “part combination kit” can be administered simultaneously, sequentially or separately. The molar ratio of the acid secretion inhibitor used according to the invention to the compound of formula I is from 1: 100 to 100: 1, such as from 1:50 to 50: 1 or from 1:20 to 20: 1, or 1 : Within the range of 10 to 10: 1. The two drugs can be administered separately at the same ratio. Examples of acid secretion inhibitors are H2 blockers such as cimetidine, ranitidine; in addition to proton pump inhibitors such as pyridinylmethylsulfinylbenzimidazoles such as omeprazole, esomeprazole, lansoprazole, pantoprazole, rabeprazole or related substances For example, reminoprazole.

Non-Medical Uses As well as compounds of formula I, salts and hydrates of such compounds, in addition to their use in therapeutic pharmaceuticals, as part of the search for new therapeutic agents, cats, dogs, rabbits, It is useful as a pharmacological tool in the development and standardization of in vitro and in vivo test systems for assessing the effects of inhibitors of activity associated with mGluR in laboratory animals such as monkeys, rats and mice.

Process for Production Another aspect of the present invention provides a process for preparing a compound of formula I or a salt or hydrate thereof. Methods for making the compounds of the present invention are described herein.

  Through the following description of such methods, appropriate protecting groups are added to various reactants and intermediates, as required, in a manner readily understood by those skilled in the art of organic synthesis, and subsequently from there. It should be understood that it is removed. Conventional techniques for using such protecting groups and examples of suitable protecting groups are described, for example, in “Protective Groups in Organic Synthesis”, TW Green, PGM Wuts, Wiley-Interscience, New York, (1999). ing. Also, conversion of a group or substituent to another group or substituent by chemical manipulation can be performed on all intermediates or final products on the synthetic route towards the final product, in which case It should be understood that this type of transformation is limited only by the inherent incompatibility of other functional groups carried on the molecule at that stage with the conditions or reagents used for the transformation. Such inherent incompatibilities and how to avoid them by performing appropriate transformation and synthesis steps in the proper order are readily understood by those skilled in the art of organic synthesis. Examples of transformations are described below, and it should be understood that the transformations described are not limited to only the general groups or substituents whose transformations are illustrated. References and explanations for other suitable transformations are described in “Comprehensive Organic Transformations-A Guide to Functional Group Preparations” R. C. Larock, VHC Publishers, Inc. (1989). References and descriptions of other suitable reactions can be found in organic chemistry texts such as “Advanced Organic Chemistry”, March, 4th ed. McGraw Hill (1992) or “Organic Synthesis”, Smith, McGraw Hill, (1994). Has been. Intermediate and final product purification techniques include, for example, normal phase and reverse phase chromatography on columns or rotating plates, recrystallization, distillation and liquid-liquid or solid-liquid extraction, and those skilled in the art. Easy to understand. The definitions of substituents and groups are as in formula I except where defined differently. The terms “room temperature” and “ambient temperature” shall mean temperatures between 16 ° C. and 25 ° C., unless otherwise specified.

  The term “reflux”, unless otherwise stated, shall mean the temperature at or above the specified solvent boiling point with respect to the solvent used.

Abbreviation
atm barometric pressure
aq. Aqueous BINAP 2,2′-bis (diphenylphosphino) -1,1′-binaphthyl Boc tert-butoxycarbonyl CDI N, N′-carbonyldiimidazole DCC N, N-dicyclohexylcarbodiimide DCM dichloromethane DBU diaza (1, 3) Bicyclo [5.4.0] undecane DEA N, N-diisopropylethylamine DIBAL-H diisobutylaluminum hydride DIC N, N'-diisopropylcarbodiimide DMAP N, N-dimethyl-4-aminopyridine DMF dimethylformamide DMSO dimethyl sulfoxide DPPF Diphenylphosphinoferrocene EA Ethyl acetate EDCI N- [3- (dimethylamino) propyl] -N′-ethylcarbodiimide hydrochloride EDC 1-ethyl-3- (3-dimethylaminopropyl) calc Diimide Et ₂ O Diethyl ether EtOAc Ethyl acetate EtOH Ethanol EtI iodoethanesulfonic ethyl ethyl Fmoc 9--fluorenylmethyloxycarbonyl h hour HetAr Heteroaryl HOBt N-hydroxybenzotriazole HBTU O-(benzotriazol-1-yl) -N, N, N ′, N′-Tetramethyluronium hexafluorophosphate HPLC High performance liquid chromatography LAH Lithium aluminum hydride LCMS HPLC Mass spectrum MCPBA m-Chlorobenzoic acid MeCN Acetonitrile MeOH Methanol
min min MeI iodomethane MeMgCl methylmagnesium chloride Me methyl n-BuLi 1-butyllithium NaOAc sodium acetate NMR nuclear magnetic resonance NMP N-methylpyrrolidinone nBuLi 1-butyllithium o.n. overnight RT, rt, rt room temperature TEA triethylamine THF tetrahydrofuran nBu Linear butyl OM mesylate or methanesulfonate ester OT tosylate, toluenesulfonate or 4-methylbenzenesulfonate ester MTBE methyl, tertbutyl ether PCC pyridinium chlorochromate PPTS pyridinium p-toluenesulfonate TBAF tetrabutylammonium fluoride pTsOH p- Toluenesulfonic acid SPE Solid phase extraction (usually silica gel for mini-chromatography No)
sat. saturated

Preparation of Intermediates Intermediates obtained by the following synthetic route are useful for the further preparation of compounds of formula I. Other starting materials are commercially available or can be prepared via methods described in the literature. The synthetic route described below is a non-limiting example of a production that can be used. Those skilled in the art understand that other routes can be used.

Synthesis of isoxazole

The aldehyde of formula VI, Scheme 1 can be used for the preparation of isoxazole. Using methods well known in the art, commercially available acid derivatives of formula II [wherein NG ¹ (G ¹ is a protecting group)] are N-protected. Thus, a compound of formula III can be obtained, wherein G ¹ is a protecting group such as Boc or. The acidic moiety in the compound of formula III can be converted to an alkyl ester of formula IV, such as a methyl or ethyl ester, which is a mild reducing agent in a solvent, such as toluene, at a low temperature, such as −78 ° C., For example, DIBAL-H can be used to convert to the aldehyde of formula VI. With higher temperatures or stronger reducing agents, primary alcohols of formula V can be formed exclusively or as a mixture with aldehydes of formula VI. Other functional groups such as primary alcohols in compounds of formula V, nitriles in compounds of formula VII, and Weinreb amide moieties in compounds of formula VIII can be prepared using formula VI, using procedures established in the art. Can be converted to aldehyde. In addition, the acid of formula II can be converted to the nitrile of formula VII by methods known in the art, for example by converting the acid to a primary amide followed by dehydration to the nitrile.

  The aldehyde of formula VI can be converted to the oxime of formula IX by treatment with hydroxylamine in a solvent such as pyridine at a temperature between 0 ° C. and room temperature. The isoxazole of formula X can be chlorinated with an oxime of formula IX using a reagent such as N-chlorosuccinimide (NCS) followed by an appropriately R-substituted acetylene derivative [where R is aryl, substituted Can be prepared by 1,3-dipolar ring addition using an aryl or masking group (eg, alkylstannane) (Steven, RV et al. Am. Chem. Soc. 1986, 108, 1039). Subsequently, isoxazole intermediate X can be deprotected by standard methods to give XI.

  Isoxazoles of formula X (wherein R is a masking group) can be prepared in this way and the masking group is converted to the desired R group by a cross-coupling reaction. For example, using a trialkylstannyl acetylene produces a trialkylstannyl isoxazole, which undergoes a reaction such as, for example, a Stille-type cross-coupling and an aryl substituent is introduced by coupling to form a suitable halogen. Can be arylated.

Synthesis of [1,2,4] -oxadiazole

The carboxylic acid of formula III can be used to prepare the corresponding 3-R substituted [1,2,4] oxadiazole of formula XII by activating the acidic moiety and the appropriate R-substituted The added hydroxyamidine forms an ester, followed by cyclization to oxadiazole. [See Tetrahedron Lett., 2001, 42, 1495-98, Tetrahedron Lett., 2001, 42, 1441-43, and Bioorg. Med. Chem. Lett. 1999, 9, 1869-74]. The acid can be activated as a mixed anhydride using an alkyl chloroformate such as isobutyl chloroformate in the presence of a base such as triethylamine in a suitable solvent such as THF. Alternatively, EDCI in a suitable solvent such as DMF, DCM, THF or MeCN, at a temperature from −20 to 100 ° C., with or without the presence of co-reagents such as HOBt or DMAP, Other well known methods for activating acids can be used, including activating acids in situ using reagents such as DCC, DIC or HBTU. Cyclization can be carried out by heating under microwave irradiation in a solvent such as pyridine or DMF, or by using a catalyst such as TBAF. R-substituted hydroxyamidines are hydroxylamine hydrochloride in a solvent such as ethanol or methanol at a temperature between room temperature and 100 ° C. in the presence of a base such as NaOH, NaHCO ₃ or Na ₂ CO _3. Is available from nitriles by adding to produce free hydroxylamine.

  A 5-R substituted [1,2,4] oxadiazole of formula XIIb is prepared from a nitrile of formula VII by effectively replacing the substituents attached to [1,2,4] -oxadiazole. can do. The nitrile of formula VII is reacted with hydroxylamine as described above to give the intermediate hydroxylamine, and an acyl containing an R group using the method described above for the conversion of a compound of formula III to a compound of formula XII. An agent may be used to convert to [1,2,4] oxadiazole of formula XIIb.

Synthesis of tetrazole

The nitrile of the formula VII is azide such as DMF, water or toluene at a temperature of 50 to 200 ° C. by conventional heating or microwave irradiation, preferably using a catalyst such as dibutyltin oxide or ZnBr _2. It can be used for the preparation of the corresponding tetrazole of formula XVIII by treatment with NaN ₃ , LiN ₃ , trialkylyltin azide or trimethylsilazide [J. Org. Chem. 2001, 7945-7950; J. Org Chem. 2000, 7984-7989 or J. Org. Chem. 1993, 4139-4141].

N2-arylation of 5-substituted tetrazoles has been reported in the literature using various coupling partners. Compounds of formula XVIII (wherein R is an aryl group) can be used as arylating agents mediated by transition metals, for example boronic acids of formula XV [having a B (OH) ₂ moiety], or the corresponding formula XVII Can be prepared using iodonium salts [with I ⁺ -Ar moiety] or the corresponding triaryl bismuth diacetates [with Bi (OAc) ₂ Ar ₂ moiety] [Tetrahedron Lett. 2002, 6221-6223; Tetrahedron Lett. 1998, 2941-2944; Tetrahedron Lett. 1999, 2747-2748]. For boronic acids, stoichiometric amounts of Cu (II) acetate and pyridine are used in solvents such as dichloromethane, DMF, dioxane or THF at temperatures from room temperature to 100 ° C. In iodonium salts, catalytic amounts of Pd (II) -compounds, such as Pd (OAc) ₂ or Pd (0) complexes, such as Pd (DBA) ₂ , or catalytic amounts of Cu (II) -carboxylates, such as Cu ( II) -Phenylcyclopropylcarboxylate and a bidentate ligand such as BINAP or DPPF are used in a solvent such as t-BuOH at a temperature of 50-100 ° C. In triaryl bismuth diacetate, a catalytic amount of cupric acetate is heated at a temperature of 40-60 ° C. in the presence of N, N, N ′, N′-tetramethylguanidine in a suitable solvent such as THF. Can be used. The iodonium salt of formula XVI is obtained, for example, from the respective boronic acid by treatment with a hypervalent iodine substituted aromatic such as hydroxyl (tosyloxy) iodobenzene or PhI (OAc) ₂ x 2TfOH in dichloromethane or the like. [See Tetrahedron Lett. 2000, 5393-5396]. Triaryl bismuth diacetate is obtained from arylmagnesium bromide using bismuth trichloride in a suitable solvent such as refluxing THF and then triaryl bismuthane and then using an oxidizing agent such as sodium perborate in acetic acid. This can be oxidized to diacetate [Synth. Commun. 1996, 4569-75].

Synthesis of amino-triazoles

The deprotected amines of formulas XI, XIII, XVIII and XIX are subjected to a series of thiourea formation, methylation and triazole formation to give a compound of formula I wherein R1 and / or R2 are as defined in formula I Can be obtained). Thiourea of formula XX can be prepared, for example, by isothiocyanate R ⁴ SCN (MeNCS as shown in Scheme 6) at a temperature between room temperature and 100 ° C. in the presence of R ² NH _{2 in} a solvent such as methanol, ethanol and the like. Or available from established methods using 1,1-thiocarbonyl-diimidazole and is typically carried out at 60 ° C. Alkylation of the thiourea intermediate is carried out in a solvent such as DMF, acetone, dichloromethane at room temperature or elevated temperature using an alkylating agent such as iodomethane (shown in Scheme 6) or iodoethane, Isothiourea of formula XXI can be obtained. When using iodoalkanes, the product can be isolated as the hydroiodide salt [see Synth. Commun. 1998, 28, 741-746]. A compound of formula XXI can be reacted with an acyl hydrazine or hydrazine followed by an acylating agent to form an intermediate that is heated at 0-150 ° C. in a suitable solvent such as pyridine or DMF. To give 3-aminotriazole of formula I.

Examples The invention will now be illustrated by the following non-limiting examples.

General methods All starting materials are commercially available or described in previous literature.
¹ H and ¹³ C NMR spectra are run on either a Bruker 300, Bruker DPX400 or Varian +400 spectrometer, for ¹ H NMR operating at 300, 400 and 400 MHz, respectively, and deuterium as solvent unless otherwise stated. Recorded in chloroformated chloroform using TMS or residual solvent signal as a reference. All reported chemical shifts are ppm on the delta-scale and are a fine splitting of the signal as shown in the record (s: singlet, brs: broad singlet, d: double Line, t: triple line, q: quadruple line, m: multiple line).

  Analysis in line liquid chromatographic separation followed by mass spectral detection was recorded on a Waters LCMS consisting of an Alliance 2795 (LC) and a ZQ integrated quadrupole mass spectrometer. The mass analyzer was equipped with an electrospray ion source operating in positive and / or negative ion mode. The ion spray voltage was ± 3 kV and the mass spectrometer was scanned from m / z 100-700 with a scan time of 0.8 seconds. Column, X-Terra MS, Waters, C8, 2.1 × 50 mm, 3.5 mm linear gradient from 5% to 100% acetonitrile in 10 mM ammonium acetate (aq) or 0.1% TFA (aq) Applied. Preparative reverse phase chromatography was run on a Gilson automated preparative HPLC with a diode array detector using XTerra MS C8, 19 x 300 mm, 7 mm as the column. Purification by chromatolone was performed on rotating silica gel / gypsum (Merck, 60 PF-254 with calcium sulfate) coated glass plates with 1, 2, or 4 mm coating layers using TC Research 7924T chromatolone. did. The product was purified by flash chromatography in a glass column packed with silica. Microwave heating was performed in a Smith Synthesizer single-mode microwave cavity that provides continuous illumination at 2450 MHz (Personal Chemistry AB, Uppsala, Sweden).

Ｎ−メチルモルホリン（９．８５ｇ，９７．５ｍｍｏｌ）及びイソブチルクロロホルメート（１３．３ｇ，９７．５ｍｍｏｌ）を−７８℃でＴＨＦ（２００ｍＬ）中の（Ｒ）−ピロリジン−１,２−ジカルボン酸１−tert−ブチルエステル（２０．０ｇ，９２．９ｍｍｏｌ）に加え、そして１時間撹拌した。反応物を室温に暖まるのにまかせながら水酸化アンモニウム（５８ｍＬ）をゆっくりと加え、そしてさらに２時間撹拌した。反応混合物をＣＨ₂Ｃｌ₂と水との間で分配し、有機抽出物を１Ｍ ＨＣｌで洗浄し、硫酸ナトリウムで乾燥させ、濾過し、そして濃縮して無色の半固形物として表題生成物（１０．８ｇ，５４％）を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 5.91−6.13 (m, 1H), 4.17−4.30 (m, 2H), 3.37−3.48 (m, 2H), 2.10−2.18 (m, 2H), 1.84−1.96 (m, 2H), 1.45 (s, 9H). Example 1: (R) -2-carbamoyl-pyrrolidine-1-carboxylic acid tert-butyl ester

N-methylmorpholine (9.85 g, 97.5 mmol) and isobutyl chloroformate (13.3 g, 97.5 mmol) were added to (R) -pyrrolidine-1,2-dicarboxylic acid in THF (200 mL) at -78 ° C. Added to 1-tert-butyl ester (20.0 g, 92.9 mmol) and stirred for 1 hour. Ammonium hydroxide (58 mL) was added slowly while allowing the reaction to warm to room temperature and stirred for an additional 2 hours. The reaction mixture is partitioned between CH ₂ Cl ₂ and water and the organic extract is washed with 1M HCl, dried over sodium sulfate, filtered and concentrated to give the title product (10 8 g, 54%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 5.91−6.13 (m, 1H), 4.17−4.30 (m, 2H), 3.37−3.48 (m, 2H), 2.10−2.18 (m, 2H) , 1.84−1.96 (m, 2H), 1.45 (s, 9H).

実施例１の表題生成物（１０．８１ｇ，５０．４５ｍｍｏｌ）及び塩化シアヌリル（５．５８ｇ，３０．３ｍｍｏｌ）をＤＭＦ（３０ｍＬ）中で１時間撹拌した。反応混合物を酢酸エチルと水との間で分配し、有機抽出物を水性炭酸ナトリウム、水、ブラインで洗浄し、硫酸ナトリウムで乾燥させ、濾過し、そして濃縮して無色の油として表題生成物（８．３４ｇ，８４％）を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 4.42−4.55 (m, 1H), 3.32−3.52 (m, 2H), 2.00−2.27 (m, 4H), 1.46−1.50 (m, 9H). Example 2: (R) -2-cyano-pyrrolidine-1-carboxylic acid tert-butyl ester

The title product of Example 1 (10.81 g, 50.45 mmol) and cyanuryl chloride (5.58 g, 30.3 mmol) were stirred in DMF (30 mL) for 1 hour. The reaction mixture was partitioned between ethyl acetate and water and the organic extract was washed with aqueous sodium carbonate, water, brine, dried over sodium sulfate, filtered and concentrated to give the title product as a colorless oil ( 8.34 g, 84%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 4.42−4.55 (m, 1H), 3.32−3.52 (m, 2H), 2.00−2.27 (m, 4H), 1.46−1.50 (m, 9H) .

実施例２の表題化合物（８．３４，４２．５ｍｍｏｌ）、アジ化ナトリウム（３．０４ｇ，４６．８ｍｍｏｌ）及び塩化アンモニウム（２．５０，４６．８ｇ）をＤＭＦ（３０ｍＬ）中、１００℃で１２時間撹拌した。反応液は濃厚物であり、そしてＤＣＭ及び３Ｍ ＨＣｌで分配した。有機抽出物を硫酸ナトリウムで乾燥し、そして濾過し、そして濃縮した。生成した固形物をエーテルで磨砕し、そして濾過して白色固形物として表題生成物（５．３１ｇ，５２％）を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 5.09−5.12 (m, 2H), 3.43−3.65 (m, 2H), 2.81−2.95 (m, 1H), 2.04−2.18 (m, 4H), 1.29−1.49 (m, 9H). Example 3: (R) -2- (2H-tetrazol-5-yl) -pyrrolidine-1-carboxylic acid tert-butyl ester

The title compound of Example 2 (8.34, 42.5 mmol), sodium azide (3.04 g, 46.8 mmol) and ammonium chloride (2.50, 46.8 g) in DMF (30 mL) at 100 ° C. Stir for 12 hours. The reaction was concentrated and partitioned with DCM and 3M HCl. The organic extract was dried over sodium sulfate, filtered and concentrated. The resulting solid was triturated with ether and filtered to give the title product (5.31 g, 52%) as a white solid.
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 5.09−5.12 (m, 2H), 3.43−3.65 (m, 2H), 2.81−2.95 (m, 1H), 2.04−2.18 (m, 4H) , 1.29-1.49 (m, 9H).

ｔ−ＢｕＯＨ（１５０ｍＬ）中の実施例３の表題化合物（４．８８ｇ，２０．４ｍｍｏｌ）、実施例１１．２の表題化合物（１１．８ｇ，２２．４ｍｍｏｌ）、ナトリウムtert−ブトキシド（２．１５ｇ，２２．４ｍｍｏｌ）、ＢＩＮＡＰ（０．５０８ｇ，０．８１６ｍｍｏｌ）、Ｐｄ₂（ＤＢＡ）₃（０．２１１ｇ，０．２０４ｍｍｏｌ）、銅２−フェニルプロパンカルボキシレート（０．１５７ｇ，０．４０８ｍｍｏｌ）を９０℃で１２時間撹拌した。反応混合物をシリカゲル上で濃縮し、そしてカラムクロマトグラフィによって精製して黄色の油として表題生成物（４．９７ｇ，６２％）を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.31 (s, 1H), 8.08 (d, 1H), 7.62 (t, 1H), 7.44 (q, 1H), 5.22−5.34 (m, 1H), 3.73−3.75 (m, 1H), 3.54−3.61 (m, 1H), 2.37−2.43 (m, 1H), 1.98−2.16 (m, 3H), 1.42 (s, 3H), 1.27 (s, 6H). Example 4.1: (R) -2- [2- (3-Bromo-phenyl) -2H-tetrazol-5-yl] -pyrrolidine-1-carboxylic acid tert-butyl ester

Title compound of Example 3 (4.88 g, 20.4 mmol), title compound of Example 11.2 (11.8 g, 22.4 mmol), sodium tert-butoxide (2.15 g) in t-BuOH (150 mL). , 22.4 mmol), BINAP (0.508 g, 0.816 mmol), Pd ₂ (DBA) ₃ (0.211 g, 0.204 mmol), copper 2-phenylpropanecarboxylate (0.157 g, 0.408 mmol). Stir at 90 ° C. for 12 hours. The reaction mixture was concentrated on silica gel and purified by column chromatography to give the title product (4.97 g, 62%) as a yellow oil.
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.31 (s, 1H), 8.08 (d, 1H), 7.62 (t, 1H), 7.44 (q, 1H), 5.22−5.34 (m, 1H ), 3.73−3.75 (m, 1H), 3.54−3.61 (m, 1H), 2.37−2.43 (m, 1H), 1.98−2.16 (m, 3H), 1.42 (s, 3H), 1.27 (s, 6H ).

In a similar manner, the following compounds were synthesized:

実施例４．１の表題化合物（４．９７ｇ，１２．６ｍｍｏｌ）、ｄｐｐｆ（０．０４２ｇ，０．０７６ｍｍｏｌ）、シアン化亜鉛（０．８９ｇ，７．５７ｍｍｏｌ）、Ｐｄ₂（ＤＢＡ）₃（０．０２６ｇ，０．０２５ｍｍｏｌ）、酢酸亜鉛（０．１８５ｇ，１．０１ｍｍｏｌ）及びＺｎ粉末（０．０６６ｇ，１．０１ｍｍｏｌ）をＤＭＦ（５０ｍＬ）及び水（１．５ｍＬ）中、９０℃で１２時間、そして１２０℃でさらに６時間撹拌した。反応混合物を酢酸エチルと水との間で分配した。有機抽出物を硫酸ナトリウムで乾燥させ、濾過し、そして濃縮し、そしてカラムクロマトグラフィによって精製して表題生成物（１．８３ｇ，４３％）を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.39−8.44 (m, 2H), 7.66−7.81 (m, 2H), 5.22−5.35 (m, 1H), 3.73−3.76 (m, 1H), 3.54−3.72 (m, 1H), 2.37−2.45 (m, 1H), 2.00−2.18 (m, 3H), 1.42 (s, 3H), 1.27 (s, 6H). Example 5: (R) -2- [2- (3-Cyano-phenyl) -2H-tetrazol-5-yl] -pyrrolidine-1-carboxylic acid tert-butyl ester

The title compound of Example 4.1 (4.97 g, 12.6 mmol), dppf (0.042 g, 0.076 mmol), zinc cyanide (0.89 g, 7.57 mmol), Pd ₂ (DBA) ₃ (0 0.026 g, 0.025 mmol), zinc acetate (0.185 g, 1.01 mmol) and Zn powder (0.066 g, 1.01 mmol) in DMF (50 mL) and water (1.5 mL) at 90 ° C. for 12 hours. And stirred at 120 ° C. for a further 6 hours. The reaction mixture was partitioned between ethyl acetate and water. The organic extract was dried over sodium sulfate, filtered and concentrated and purified by column chromatography to give the title product (1.83 g, 43%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.39−8.44 (m, 2H), 7.66−7.81 (m, 2H), 5.22−5.35 (m, 1H), 3.73−3.76 (m, 1H) , 3.54−3.72 (m, 1H), 2.37−2.45 (m, 1H), 2.00−2.18 (m, 3H), 1.42 (s, 3H), 1.27 (s, 6H).

ＤＣＭ（６．０ｍＬ）中の実施例５の表題化合物（０．９１９ｇ，２．７０ｍｍｏｌ）を０℃でＴＦＡ（６ｍＬ）に加えた。反応液を３０分撹拌し、そして濃縮した。それをＤＣＭ及び２Ｍ Ｎａ₂ＣＯ₃で分配した。有機抽出物を硫酸ナトリウムで乾燥させ、濾過し、そして濃縮して薄黄色の固形物（０．５９３ｇ，９２％）として表題生成物を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.47 (t, 1H), 8.43 (dd, 1H), 7.78 (dd, 1H), 7.13
(t, 1H), 4.66 (q, 1H), 3.23−3.25 (m, 1H), 3.12−3.21 (m, 1H), 2.20−2.42 (m, 2H), 2.12−2.19 (m, 2H), 1.96−2.04 (m, 2H). Example 6.1: 3-((R) -5-pyrrolidin-2-yl-tetrazol-2-yl) -benzonitrile

The title compound of Example 5 (0.919 g, 2.70 mmol) in DCM (6.0 mL) was added to TFA (6 mL) at 0 ° C. The reaction was stirred for 30 minutes and concentrated. It was partitioned with DCM and 2M Na ₂ CO ₃ . The organic extract was dried over sodium sulfate, filtered and concentrated to give the title product as a pale yellow solid (0.593 g, 92%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.47 (t, 1H), 8.43 (dd, 1H), 7.78 (dd, 1H), 7.13
(t, 1H), 4.66 (q, 1H), 3.23−3.25 (m, 1H), 3.12−3.21 (m, 1H), 2.20−2.42 (m, 2H), 2.12−2.19 (m, 2H), 1.96 −2.04 (m, 2H).

In a similar manner, the following compounds were synthesized:

ＣＨ₃Ｃｌ（１０ｍＬ）中の実施例６．１の表題化合物（０．５９３ｇ，２．４７ｍｍｏｌ）及び（メチルイミノ）（チオキソ）メタン（０．２８７ｇ，３．９３ｍｍｏｌ）を１．５時間撹拌した。反応混合物を濃縮し、そしてエーテル中で磨砕し、濾過してオフホワイトの固形物として表題生成物を得た（０．６５６ｇ，８５％）。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.39−8.44 (m, 2H), 7.78 (dd, 1H), 7.72 (t, 1H),
5.89−5.99 (m, 2H), 3.68−3.77 (m, 1H), 3.46−3.53 (m, 1H), 3.15 (d, 3H), 2.38−2.45 (m, 2H), 2.24−2.26 (m, 2H). Example 7.1: (R) -2- [2- (3-cyano-phenyl) -2H-tetrazol-5-yl] -pyrrolidine-1-carbothioic acid methylamide

The title compound of Example 6.1 (0.593 g, 2.47 mmol) and (methylimino) (thioxo) methane (0.287 g, 3.93 mmol) in CH ₃ Cl (10 mL) were stirred for 1.5 hours. The reaction mixture was concentrated and triturated in ether and filtered to give the title product as an off-white solid (0.656 g, 85%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.39-8.44 (m, 2H), 7.78 (dd, 1H), 7.72 (t, 1H),
5.89−5.99 (m, 2H), 3.68−3.77 (m, 1H), 3.46−3.53 (m, 1H), 3.15 (d, 3H), 2.38−2.45 (m, 2H), 2.24−2.26 (m, 2H ).

In a similar manner, the following compounds were synthesized:

ＭｅＯＨ（５．０ｍＬ）中の実施例７．２の表題化合物（０．３８５ｇ，１．２０ｍｍｏｌ）及びヨウ化メチル（０．２９８ｇ，２．１ｍｍｏｌ）を８０℃で１時間撹拌した。反応液を濃縮し、ＤＣＭ及び炭酸ナトリウムで分配した。有機抽出物をブラインで洗浄し、硫酸ナトリウムで乾燥させ、濾過し、そして濃縮して琥珀色の油として表題生成物（０．４００ｇ，８８％）を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.15 (t, 1H), 8.03 (dt, 1H), 7.43−7.51 (m, 2H),
5.60−5.63 (m, 1H), 3.82−3.84 (m, 1H), 3.67−3.70 (m, 1H), 3.19 (s, 3H), 2.40−2.43 (m, 1H), 2.27 (s, 3H), 2.02−2.17 (m, 3H). Example 8: (R) -2- [2- (3-Chloro-phenyl) -2H-tetrazol-5-yl] -N-methyl-pyrrolidine-1-carboxymidothioic acid methyl ester

The title compound of Example 7.2 (0.385 g, 1.20 mmol) and methyl iodide (0.298 g, 2.1 mmol) in MeOH (5.0 mL) were stirred at 80 ° C. for 1 hour. The reaction was concentrated and partitioned with DCM and sodium carbonate. The organic extract was washed with brine, dried over sodium sulfate, filtered and concentrated to give the title product (0.400 g, 88%) as an amber oil.
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.15 (t, 1H), 8.03 (dt, 1H), 7.43-7.51 (m, 2H),
5.60−5.63 (m, 1H), 3.82−3.84 (m, 1H), 3.67−3.70 (m, 1H), 3.19 (s, 3H), 2.40−2.43 (m, 1H), 2.27 (s, 3H), 2.02−2.17 (m, 3H).

実施例７．１の表題化合物（０．６５６ｇ，２．０９ｌ）及びナトリウムtert−ブトキシド（０．２０１，２．０９ｍｍｏｌ）をＴＨＦ（５ｍＬ）中で撹拌した。ＴＨＦ（２ｍＬ）中のヨウ化メチル（０．８９ｇ，０．６２４ｍｍｏｌ）を加え、そして反応液を２０分間撹拌した。反応混合物を水中へ注ぎ、そして酢酸エチルで分配した。有機抽出物を水、ブラインで洗浄し、硫酸ナトリウムで乾燥させ、濾過し、そして濃縮して琥珀色の油として表題生成物を得た（０．５２２ｇ，７６％）。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.43− 8.39 (m, 2H), 7.76 (dd, 1H), 7.70 (t, 1H), 5.59−5.63 (m, 1H), 3.83−3.85 (m, 1H), 3.68−3.71(m, 1H), 2.40−2.51 (m, 1H), 2.27 (s, 3H), 2.06−2.17 (m, 3H). Example 9: (R) -2- [2- (3-Cyano-phenyl) -2H-tetrazol-5-yl] -N-methyl-pyrrolidine-1-carboxymidothioic acid methyl ester

The title compound of Example 7.1 (0.656 g, 2.09 l) and sodium tert-butoxide (0.201, 2.09 mmol) were stirred in THF (5 mL). Methyl iodide (0.89 g, 0.624 mmol) in THF (2 mL) was added and the reaction was stirred for 20 minutes. The reaction mixture was poured into water and partitioned with ethyl acetate. The organic extract was washed with water, brine, dried over sodium sulfate, filtered and concentrated to give the title product as an amber oil (0.522 g, 76%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.43− 8.39 (m, 2H), 7.76 (dd, 1H), 7.70 (t, 1H), 5.59−5.63 (m, 1H), 3.83−3.85 (m, 1H), 3.68-3.71 (m, 1H), 2.40-2.51 (m, 1H), 2.27 (s, 3H), 2.06-2.17 (m, 3H).

１−クロロ−３−ヨードベンゼン（５．０ｇ，２１ｍｍｏｌ）を３０℃で撹拌した。過酢酸（４０％，８．３５ｍＬ，５０．３ｍｍｏｌ）を溶液に滴加し、そして反応液を１２時間撹拌した。形成された白色固形物を濾過し、１０％酢酸で１回及びヘキサンで３回洗浄し、そして真空で乾燥して白色固形物として表題生成物（２７．５ｇ，９２％）を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.10 (s, 1H), 7.99 (d, 1H), 7.57 (d, 1H), 7.46 (t, 1H), 2.04 (s, 6H). Example 10.1 m-chlorophenyl iodine diacetate

1-Chloro-3-iodobenzene (5.0 g, 21 mmol) was stirred at 30 ° C. Peracetic acid (40%, 8.35 mL, 50.3 mmol) was added dropwise to the solution and the reaction was stirred for 12 hours. The white solid formed was filtered, washed once with 10% acetic acid and three times with hexane and dried in vacuo to give the title product (27.5 g, 92%) as a white solid.
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.10 (s, 1H), 7.99 (d, 1H), 7.57 (d, 1H), 7.46 (t, 1H), 2.04 (s, 6H).

In a similar manner, the following compounds were synthesized:

三フッ化ホウ素ジエチルエーテラート（１６．５１ｇ，１１６．３ｍｍｏｌ）を−５℃でＤＣＭ（１７０ｍｌ）中の３−クロロフェニルボロン酸（１７．３７ｇ，１１１．０ｍｍｏｌ）に撹拌しながらゆっくりと加えた。１５分後、ＤＣＭ（１５０ｍＬ）中の実施例１０．１の表題化合物（３７．７１ｇ，１０５．８ｍｍｏｌ）をゆっくりと加えた。反応液を０℃で１時間撹拌し、そしてテトラフルオロホウ酸ナトリウム（水３００ｍＬ中２２５ｇ）を加え、そして１時間撹拌した。有機層を分離し、硫酸ナトリウムで乾燥させ、濾過し、そして濃縮し、そしてエーテルで磨砕して薄茶色の固形物として表題生成物（３１．６ｇ，６８％）を得た。
¹H NMR (300 MHz, (CD₃)₂SO):δ(ppm) 7.60 (t, 2H), 7.74 (dd, 2H), 8.26 (dd, 2H), 8.50 (s, 2H). Example 11.1 Bis (3-chlorophenyl) iodonium tetrafluoroborate

Boron trifluoride diethyl etherate (16.51 g, 116.3 mmol) was slowly added with stirring to 3-chlorophenylboronic acid (17.37 g, 111.0 mmol) in DCM (170 ml) at −5 ° C. After 15 minutes, the title compound of Example 10.1 (37.71 g, 105.8 mmol) in DCM (150 mL) was added slowly. The reaction was stirred at 0 ° C. for 1 hour and sodium tetrafluoroborate (225 g in 300 mL of water) was added and stirred for 1 hour. The organic layer was separated, dried over sodium sulfate, filtered and concentrated and triturated with ether to give the title product (31.6 g, 68%) as a light brown solid.
¹ H NMR (300 MHz, (CD ₃ ) ₂ SO): δ (ppm) 7.60 (t, 2H), 7.74 (dd, 2H), 8.26 (dd, 2H), 8.50 (s, 2H).

In a similar manner, the following compounds were synthesized:

ＤＭＦ（２２０ｍＬ）中の２−クロロ−６−メトキシ−イソニコチン酸（１６ｇ，８５ｍｍｏｌ）にＫ₂ＣＯ₃（４７ｇ，３４１ｍｍｏｌ）及びＭｅＩ（６．３７ｍＬ，１０２ｍｍｏｌ）を加えた。一夜撹拌した後、反応混合物を濾過し、それから濃縮した。残留物を酢酸エチル中に溶解し、水（３回）及びブラインで洗浄し、無水Ｎａ₂ＳＯ₄で乾燥させ、濾過し、そして濃縮した。ヘキサン中１０−３０％酢酸エチルで溶出するフラッシュカラムクロマトグラフィにより精製して表題生成物（１５ｇ，８７％）を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 7.45 (s, 1H), 7.23 (s, 1H), 3.98 (s, 3H), 3.95 (s, 3H). Example 12.1: 2-Chloro-6-methoxy-isonicotinic acid methyl ester

DMF (220 mL) of 2-chloro-6-methoxy - isonicotinic acid (16g, 85 mmol) in _{K 2 CO 3 (47g, 341mmol} ) and MeI (6.37mL, 102mmol) was added. After stirring overnight, the reaction mixture was filtered and then concentrated. The residue was dissolved in ethyl acetate, washed with water (3 times) and brine, dried over anhydrous Na ₂ SO ₄ , filtered and concentrated. Purification by flash column chromatography eluting with 10-30% ethyl acetate in hexanes afforded the title product (15 g, 87%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 7.45 (s, 1H), 7.23 (s, 1H), 3.98 (s, 3H), 3.95 (s, 3H).

In a similar manner, the following compounds were synthesized:

実施例１２．１の表題化合物（１５ｇ，７４．８ｍｍｏｌ）をエタノール（３５０ｍＬ）中のＰｄ／Ｃ（７．４ｇ，８２．２ｍｍｏｌ）と混合した。
反応混合物をフラッシュし、そして水素を充填し、それから室温で一夜撹拌した。Celite^(R)パッドを通して反応混合物を濾過し、そして真空で濃縮した。残留物をジクロロメタンに溶解し、そして水で２回及びブラインで洗浄した。有機相を無水硫酸ナトリウムで乾燥させ、濾過し、そして真空で濃縮して生成物（９．５１ｇ，７５％）として薄黄色の油を得た。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.29 (d, 1H), 7.41 (d, 1H), 7.32 (s, 1H), 3.98 (s, 3H), 3.95 (s, 3H). Example 13.1: 2-Methoxy-isonicotinic acid methyl ester

The title compound of Example 12.1 (15 g, 74.8 mmol) was mixed with Pd / C (7.4 g, 82.2 mmol) in ethanol (350 mL).
The reaction mixture was flushed and charged with hydrogen and then stirred at room temperature overnight. The reaction mixture was filtered through Celite ^(R) pad, and concentrated in vacuo. The residue was dissolved in dichloromethane and washed twice with water and brine. The organic phase was dried over anhydrous sodium sulfate, filtered and concentrated in vacuo to give a pale yellow oil as the product (9.51 g, 75%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.29 (d, 1H), 7.41 (d, 1H), 7.32 (s, 1H), 3.98 (s, 3H), 3.95 (s, 3H).

In a similar manner, the following compounds were synthesized:

エタノール（１００ｍＬ）中の実施例１３．１の表題化合物（９．５１ｍｇ，５７．０ｍｍｏｌ）にヒドラジン水和物（３．４５ｍＬ，７１．２ｍｍｏｌ）を加え、それから７８℃で一夜加熱した。反応混合物を冷却し、そして真空で濃縮した。残留物を酢酸エチルで磨砕し、濾過し、そして乾燥して白色の固形物として表題生成物を得た（６．６９ｍｇ，７０．３％）。
¹H NMR (300 MHz, (CD₃)₂SO):δ(ppm) 10.04 (br, 1H), 8.27 (d, 1H), 7.32 (d, 1H), 7.15 (s, 1H), 4.62 (br, 2H), 3.88 (s, 3H). Example 14.1: 2-methoxy-isonicotinic hydrazide

To the title compound of Example 13.1 (9.51 mg, 57.0 mmol) in ethanol (100 mL) was added hydrazine hydrate (3.45 mL, 71.2 mmol) and then heated at 78 ° C. overnight. The reaction mixture was cooled and concentrated in vacuo. The residue was triturated with ethyl acetate, filtered and dried to give the title product as a white solid (6.69 mg, 70.3%).
¹ H NMR (300 MHz, (CD ₃ ) ₂ SO): δ (ppm) 10.04 (br, 1H), 8.27 (d, 1H), 7.32 (d, 1H), 7.15 (s, 1H), 4.62 (br , 2H), 3.88 (s, 3H).

In a similar manner, the following compounds were synthesized:

実施例８の表題化合物（６０ｍｇ，０．１８ｍｍｏｌ）及び実施例１４．２の表題生成物（５３ｍｇ，０．３５ｍｍｏｌ）をイソプロパノール（２ｍＬ）中で混合し、撹拌棒付きのバイアル中に置き、そして乾式冷却器を備えつけた。反応混合物を９０℃で２４時間撹拌した。反応混合物を濃縮し、それからＤＣＭで希釈した。ポリマーに支持されたイソシアネートを加え、そして混合物を撹拌して過剰の２−メチルイソニコチノヒドラジドを除去した。混合物を濾過し、そして濾液を濃縮した。粗残留物をエーテルで一夜磨砕した。磨砕して表題生成物として淡黄色の固形物が形成された（３５ｍｇ，４７％）。
¹H NMR (300 MHz, CDCl₃):δ(ppm) 8.61 (m, 1H), 8.12 (m, 1H), 8.08 (m, 1H), 7.58 (m, 1H), 7.46 (m, 3H), 5.75 (m, 1H), 4.00 (m, 1H), 3.69 (s, 3H), 3.57 (m, 1H), 2.70 (s, 3H), 2.36 (m, 4H). Example 15.1: 4- (5-{(R) -2- [2- (3-chloro-phenyl) -2H-tetrazol-5-yl] -pyrrolidin-1-yl} -4-methyl-4H -[1,2,4] triazol-3-yl) -2-methyl-pyridine

The title compound of Example 8 (60 mg, 0.18 mmol) and the title product of Example 14.2 (53 mg, 0.35 mmol) are mixed in isopropanol (2 mL), placed in a vial with a stir bar, and A dry cooler was installed. The reaction mixture was stirred at 90 ° C. for 24 hours. The reaction mixture was concentrated and then diluted with DCM. The polymer supported isocyanate was added and the mixture was stirred to remove excess 2-methylisonicotinohydrazide. The mixture was filtered and the filtrate was concentrated. The crude residue was triturated with ether overnight. Trituration formed a pale yellow solid as the title product (35 mg, 47%).
¹ H NMR (300 MHz, CDCl ₃ ): δ (ppm) 8.61 (m, 1H), 8.12 (m, 1H), 8.08 (m, 1H), 7.58 (m, 1H), 7.46 (m, 3H), 5.75 (m, 1H), 4.00 (m, 1H), 3.69 (s, 3H), 3.57 (m, 1H), 2.70 (s, 3H), 2.36 (m, 4H).

In a similar manner, the following compounds were synthesized:

Biological Evaluation Functional Evaluation of mGluR5 Antagonism in Cell Lines Expressing mGluR5D The properties of the compounds of the invention can be analyzed using standard assays for pharmacological activity. Examples of glutamate receptor assays are described in, for example, Aramori et al., Neuron 8: 757 (1992), Tanabe et al., Neuron 8: 169 (1992), Miller et al., J. Neuroscience 15: 6103 (1995), It is well known in the art as described by Balazs, et al., J. Neurochemistry 69: 151 (1997). The methodologies described in these publications are incorporated herein by reference. Conveniently, the compounds of the present invention may be used in assays that measure the mobilization of intracellular calcium [Ca ²⁺ ] _i in cells expressing mGluR5 (FLIPR), or in other assays that measure inositol phosphate turnover ( Can be studied by IP3).

FLIPR assay Cells expressing human mGluR5d as described in WO 97/05252 are seeded at a density of 100,000 cells per well on a collagen-coated transparent bottom 96-well plate with black sides, and Experiments were performed 24 hours after sowing. All assays consisted of 127 mM NaCl, 5 mM KCl, 2 mM MgCl2, 0.7 mM NaH ₂ PO ₄ , 2 mM CaCl ₂ , 0.422 mg / mL NaHCO ₃ , 2.4 mg / mL HEPES, 1.8 mg / mL glucose and 1 mg / mL It was carried out in a buffer containing mL BSA fraction IV (pH 7.4). Cell cultures in 96-well plates were placed in 0.01% pluronic acid (registered trademark nonionic activator polyol-CAS No. 9003-11-6) with fluorescent calcium indicator Fluo-3 (Molecular Probes, Eugene, Oregon). ) In the above buffer containing 4 μM acetoxymethyl ester type. After the loading period, the fluo-3 buffer was removed and replaced with fresh assay buffer. The FLIPR experiment was conducted using excitation and emission wavelengths of 488 nm and 562 nm, respectively, with a laser setting of 0.800 W and a CCD camera shutter speed of 0.4 seconds. Each experiment started with 160 μl of buffer in each well of the cell plate. After adding 40 μl from the antagonist plate, 50 μL was added from the agonist plate. Antagonist and agonist additions were separated by 90 seconds. The fluorescence signal was taken 50 times at 1 second intervals, followed by 3 samples at 5 second intervals immediately after each addition of 2 each. Response was measured as the difference between the peak height of the response to the agonist and the lower background fluorescence within the sampling period. IC ₅₀ measurements were made using a linear least squares fitting program.

IP3 Assay A further functional assay for mGluR5d is described in WO 97/05252 and is based on phosphatidylinositol turnover. Receptor activation stimulates phospholipase C activity and increases the formation of inositol 1,4,5 triphosphate (IP ₃ ). GHEK, which stably expresses human mGluR5d, was seeded onto a 24-well poly-L-lysine-coated plate at 40 × 10 ⁴ cells / well in a medium containing 1 μCi / well [3H] myo-inositol. Cells were incubated overnight (16 hours), then washed 3 times and HEPES buffered saline supplemented with 1 unit / ml glutamate pyruvate transaminase and 2 mM pyruvate (146 mM NaCl, 4.2 mM KCl, 0.5 mM MgCl ₂ , 0.1% glucose, 20 mM HEPES, pH 7.4) and incubated at 37 ° C. for 1 hour. Cells were washed once in HEPES buffered saline and preincubated for 10 minutes in HEPES buffered saline containing 10 mM LiCl. Compounds were incubated in duplicate for 15 minutes at 37 ° C., then glutamic acid (80 μM) or DHPG (30 μM) was added and incubated for an additional 30 minutes. The reaction was terminated by adding 0.5 mL of perchloric acid (5%) on ice and incubated at 4 ° C. for at least 30 minutes. Samples were collected in 15 mL polypropylene tubes and inositol phosphates were separated using ion exchange resin (Dowex AG1-X8 formate form, 200-400 mesh, BIORAD) columns. Separation of inositol phosphate was performed by first eluting glycerophosphatidylinositol with 8 mL of 30 mM ammonium formate. Total inositol phosphate was then eluted with 8 mL of 700 mM ammonium formate / 100 mM formic acid and collected in scintillation vials. The eluate was then mixed with 8 mL of scintillant and [3H] inositol incorporation was measured by scintillation counting. The dpm counts were plotted from duplicate samples and IC ₅₀ measurements were obtained using a linear least squares fitting program.

Abbreviation BSA Bovine Serum Albumin CCD Charge Coupled Device CRC Concentration Response Curve DHPG 3,5-Dihydroxyphenylglycine DPM Number of decays per minute EDTA Ethylenediaminetetraacetic acid FLIPR Fluorescent quantitative imaging plate reader GHEK GLAST included GLAST Transporter HEPES 4- (2-hydroxyethyl) -1-piperazineethanesulfonic acid (buffer)
IP ₃ inositol triphosphate

In general, in the above assay, compounds with IC ₅₀ values less than 10000 nM were active. In one embodiment of the invention, the IC ₅₀ value is less than 1000 nM. In a further aspect of the invention, the IC ₅₀ value is less than 100 nM.

Measurement of brain-to-plasma ratio in rats Brain-to-plasma ratio was assessed in female Sprague Dawley rats. The compound was dissolved in water or another suitable vehicle. In determining the brain-to-plasma ratio, the compound was administered as a subcutaneous or intravenous bolus injection, or intravenous infusion, or oral administration. Blood samples were collected by cardiac puncture at predetermined time points after administration. Rats were killed by incising the heart and the brain was immediately retained. To separate plasma from blood cells, blood samples were collected in heparinized tubes and centrifuged within 30 minutes. Plasma was transferred to 96 well plates and stored at -20 ° C until analysis. The brain was divided in half and each half was placed in a pre-tarred tube and stored at −20 ° C. until analysis. Prior to analysis, brain samples were thawed and 3 mL / g brain tissue of distilled water was added to the tubes. Brain samples were sonicated in an ice bath until the samples were uniform. Both brain and plasma samples were precipitated with acetonitrile. After centrifugation, the supernatant was diluted with 0.2% formic acid. The analysis was performed on a short path reverse phase HPLC column with rapid gradient elution and MSMS detection using a triple quadrupole instrument with electrospray ionization and Selected Reaction Monitoring acquisition. Liquid-liquid extraction can be used as an alternative to sample cleaning. After adding the appropriate buffer, the sample is extracted into an organic solvent by shaking. An aliquot of the organic layer was transferred to a new vial and evaporated to dryness under a stream of nitrogen. After reconstituting the residue, the sample was ready for injection onto the HPLC column.

  In general, the compounds of the present invention are constrained in extension with a drug ratio in plasma / drug in the brain of <0.5 in rats. In one embodiment, the ratio is less than 0.15.

Measurement of in vitro stability Rat liver microsomes were prepared from Sprague-Dawley rat liver samples. Human liver microsomes were prepared from human liver samples or obtained from BD Gentest. Compounds were incubated at 37 ° C. at a total microsomal protein concentration of 0.5 mg / ml in the presence of the cofactor NADPH (1.0 mmol / l) in 0.1 mol / L potassium phosphate buffer at pH 7.4. The initial concentration of the compound was 1.0 μmol / L. For analysis, samples were taken at 5 time points 0, 7, 15, 20 and 30 minutes after the start of incubation. Enzyme activity in the collected sample was immediately stopped by adding 3.5 volumes of acetonitrile. The concentration of compound remaining in each of the collected samples was determined by LC-MS. The elimination rate constant (k) of the mGluR5 inhibitor was calculated as the slope of the plot of In [mGluR5 inhibitor] versus incubation time (minutes). The elimination rate constant is then used to calculate the half-life (T 1/2) of the mGluR5 inhibitor, which is then used to determine the intrinsic clearance (CLint) of the mGluR5 inhibitor in liver microsomes:
CLint. = (Ln2 × incubation volume) / (T 1/2 xprotein concentration) = μl / min / mg
Calculated as

Screening for compounds active against TLESR An adult Labrador retriever of both sexes trained to stand in the Pavlov Sling was used. Esophageal fistula formation from the mucosa to the skin was performed and the dog was fully recovered before all experiments were performed.

Motility measurement In summary, after free feeding for about 17 hours with free supply of water, a multi lumen sleeve / sidehole assembly (Dentsleeve, Adelaide, South Australia) was used for esophageal fistula formation. The gastric lower esophageal sphincter (LES) and esophageal pressure were measured. The assembly was perfused with water using a low compliance pressure gauge infusion pump (Dentsleeve, Adelaide, South Australia). Swallowing was measured by passing through a tube perfused with air in the oral direction, and the pH was monitored with an antimony electrode 3 cm above the LES. All signals were amplified and obtained on a personal computer at 10 Hz.

  Placebo (0.9% NaCl) or test compound was administered intravenously in the forelimb vein when a baseline measurement without fasting stomach / LES phase 3 motor activity was obtained (iv, 0 .5 mL / kg). Ten minutes after intravenous administration, a nutrient diet (10% peptone, 5% D-glucose, 5% intralipid, pH 3.0) was infused into the stomach at 100 mL / min through the central lumen of the assembly to a final volume of 30 mL / kg. . Following the infusion of the nutrient meal, air was infused at a rate of 500 mL / min until an intragastric pressure of 10 ± 1 mmHg was obtained. The pressure was then maintained at this level during the experiment using an infusion pump to further inflate or degas air from the stomach. The experimental time from the start of nutrient infusion to the end of air aeration was 45 minutes. The approach was effective as a reliable means of inducing TLESR.

  TLESR is defined as the decrease in lower esophageal sphincter pressure (relative to intragastric pressure) at a rate of> 1 mmHg / sec. If relaxation is classified as swallow-induced, no pharyngeal signal should occur ≦ 2 seconds before the occurrence of relaxation. The pressure difference between the LES and the stomach must be less than 2 mm Hg and the duration of complete relaxation must be longer than 1 second.

The sample results are shown in the following table:

Claims (27)

Formula (I)

(Where
R ¹ is methyl, halogen or cyano;
R ² is hydrogen or fluoro;
R ³ is hydrogen, fluoro or C ₁ -C ₃ alkyl;
R ⁴ is C ₁ -C ₃ alkyl or cyclopropyl;
X is

Is;
And Z is

Is;
put it here,
R ⁵ is hydrogen, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, C ₁ -C ₃ alkoxy; C ₁ -C ₃ haloalkoxy; or halogen;
R ⁶ is hydrogen, C ₁ -C ₃ alkyl, C ₁ -C ₃ haloalkyl, C ₁ -C ₃ alkoxy; C ₁ -C ₃ haloalkoxy; or halogen;
R ⁷ is hydrogen, fluoro or C ₁ -C ₃ alkyl)
And pharmaceutically acceptable salts, hydrates, isoforms, tautomers and / or enantiomers thereof. The compound according to claim 1, wherein R ¹ is halogen or cyano. 3. A compound according to claim 2, wherein R < ¹ > is chloro. The compound of claim 2, wherein R ¹ is cyano. The compound according to any one of claims 1 to 4, wherein R ² is hydrogen. R ³ is hydrogen or fluoro, A compound according to any one of claims 1 to 5. R ⁴ is C ₁ -C ₂ alkyl A compound according to any one of claims 1-6. R ⁴ is methyl A compound according to claim 7. R ⁵ is hydrogen, C ₁ -C ₂ alkyl or C ₁ -C ₂ alkoxy, A compound according to any one of claims 1-8. R ⁶ is hydrogen, C ₁ -C ₂ alkyl or C ₁ -C ₂ alkoxy, A compound according to any one of claims 1 to 9. R ⁷ is C ₁ -C ₂ alkyl or C ₁ -C ₂ alkoxy, A compound according to any one of claims 1 to 10. 4- (5-{(R) -2- [2- (3-Chloro-phenyl) -2H-tetrazol-5-yl] -pyrrolidin-1-yl} -4-methyl-4H- [1,2, 4] triazol-3-yl) -2-methyl-pyridine;
3- {5-[(R) -1- (4-Methyl-5-pyridin-3-yl-4H- [1,2,4] triazol-3-yl) -pyrrolidin-2-yl] -tetrazole- 2-yl} -benzonitrile;
3- (5-{(R) -1- [5- (2-methoxy-pyridin-4-yl) -4-methyl-4H- [1,2,4] triazol-3-yl] -pyrrolidine-2 -Yl} -tetrazol-2-yl) -benzonitrile; and 3- (5-{(R) -1- [4-methyl-5- (2-methyl-pyridin-4-yl) -4H- [1 , 2,4] triazol-3-yl] -pyrrolidin-2-yl} -tetrazol-2-yl) -benzonitrile and pharmaceutically acceptable salts, hydrates, isoforms, tautomers thereof Sex and / or enantiomer.   13. A compound according to any one of claims 1 to 12 for use in therapy.   A pharmaceutical composition comprising the compound according to any one of claims 1 to 12 as an active ingredient together with a pharmacologically and pharmaceutically acceptable carrier.   Use of the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt or optical isomer thereof for the manufacture of a medicament that inhibits transient lower esophageal sphincter relaxation.   Use of the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt or optical isomer thereof for the manufacture of a medicament for treating or preventing gastroesophageal reflux disease.   Use of the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt or optical isomer thereof for the manufacture of a medicament for treating or preventing pain.   Use of the compound according to any one of claims 1 to 12, or a pharmaceutically acceptable salt or optical isomer thereof for the manufacture of a medicament for treating or preventing anxiety.   Use of the compound according to any one of claims 1 to 12 or a pharmaceutically acceptable salt or optical isomer thereof for the manufacture of a medicament for treating or preventing irritable bowel syndrome (IBS).   13. A method of inhibiting transient lower esophageal sphincter relaxation by administering an effective amount of a compound according to any one of claims 1-12 to a subject in need of such inhibition.   A method of treating or preventing gastroesophageal reflux disease, comprising administering an effective amount of a compound according to any one of claims 1 to 12 to a subject in need of such treatment or prevention. Method.   A method for treating or preventing pain, comprising administering an effective amount of a compound according to any one of claims 1 to 12 to a subject in need of such treatment or prevention.   A method of treating or preventing anxiety, comprising administering an effective amount of a compound according to any one of claims 1 to 12 to a subject in need of such treatment or prevention.   A method of treating or preventing irritable bowel syndrome (IBS), comprising administering an effective amount of the compound according to any one of claims 1 to 12 to a subject in need of such treatment or prevention. According to the method. A combination comprising (i) at least one compound according to any one of claims 1 to 12, and (ii) at least one acid secretion inhibitor.   26. The combination according to claim 25, wherein the acid secretion inhibitor is selected from cimetidine, ranitidine, omeprazole, esomeprazole, lansoprazole, pantoprazole, rabeprazole or reminoprazole. (R) -2- (2H-tetrazol-5-yl) -pyrrolidine-1-carboxylic acid tert-butyl ester;
(R) -2- [2- (3-Bromo-phenyl) -2H-tetrazol-5-yl] -pyrrolidine-1-carboxylic acid tert-butyl ester;
(R) -2- [2- (3-Chloro-phenyl) -2H-tetrazol-5-yl] -pyrrolidine-1-carboxylic acid tert-butyl ester;
(R) -2- [2- (3-Cyano-phenyl) -2H-tetrazol-5-yl] -pyrrolidine-1-carboxylic acid tert-butyl ester;
3-((R) -5-pyrrolidin-2-yl-tetrazol-2-yl) -benzonitrile;
2- (3-chloro-phenyl) -5- (R) -pyrrolidin-2-yl-2H-tetrazole;
(R) -2- [2- (3-Cyano-phenyl) -2H-tetrazol-5-yl] -pyrrolidine-1-carbothioic acid methylamide;
(R) -2- [2- (3-Chloro-phenyl) -2H-tetrazol-5-yl] -pyrrolidine-1-carbothioic acid methylamide;
(R) -2- [2- (3-Chloro-phenyl) -2H-tetrazol-5-yl] -N-methyl-pyrrolidine-1-carboximidothioic acid methyl ester; and (R) -2- [2 A compound selected from-(3-cyano-phenyl) -2H-tetrazol-5-yl] -N-methyl-pyrrolidine-1-carboxymidothioic acid methyl ester.